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. 1990 Jun;172(6):3490–3495. doi: 10.1128/jb.172.6.3490-3495.1990

Cloning and sequence analysis of tryptophan synthetase genes of an extreme thermophile, Thermus thermophilus HB27: plasmid transfer from replica-plated Escherichia coli recombinant colonies to competent T. thermophilus cells.

Y Koyama 1, K Furukawa 1
PMCID: PMC209164  PMID: 2188962

Abstract

Tryptophan synthetase genes (trpBA) of the extreme thermophile Thermus thermophilus HB27 were cloned by a novel method of direct plasmid transfer from replica-plated Escherichia coli recombinant colonies to competent T. thermophilus HB27 trpB cells. The nucleotide sequences of the trpBA genes were determined. The amino acid sequences deduced from the nucleotide sequences of Thermus trpB and trpA were found to have identities of 54.8 and 28.7%, respectively, with those of E. coli trpB and trpA genes. Low cysteine content (one in trpB; zero in trpA) is a striking feature of these proteins, which may contribute to their thermostability.

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Selected References

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  1. Aksoy S., Squires C. L., Squires C. Translational coupling of the trpB and trpA genes in the Escherichia coli tryptophan operon. J Bacteriol. 1984 Feb;157(2):363–367. doi: 10.1128/jb.157.2.363-367.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  3. Chang M., Hadero A., Crawford I. P. Sequence of the Pseudomonas aeruginosa trpI activator gene and relatedness of trpI to other procaryotic regulatory genes. J Bacteriol. 1989 Jan;171(1):172–183. doi: 10.1128/jb.171.1.172-183.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crawford I. P., Nichols B. P., Yanofsky C. Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium. J Mol Biol. 1980 Oct 5;142(4):489–502. doi: 10.1016/0022-2836(80)90259-4. [DOI] [PubMed] [Google Scholar]
  5. Hadero A., Crawford I. P. Nucleotide sequence of the genes for tryptophan synthase in Pseudomonas aeruginosa. Mol Biol Evol. 1986 May;3(3):191–204. doi: 10.1093/oxfordjournals.molbev.a040388. [DOI] [PubMed] [Google Scholar]
  6. Hyde C. C., Ahmed S. A., Padlan E. A., Miles E. W., Davies D. R. Three-dimensional structure of the tryptophan synthase alpha 2 beta 2 multienzyme complex from Salmonella typhimurium. J Biol Chem. 1988 Nov 25;263(33):17857–17871. [PubMed] [Google Scholar]
  7. Kagawa Y., Nojima H., Nukiwa N., Ishizuka M., Nakajima T., Yasuhara T., Tanaka T., Oshima T. High guanine plus cytosine content in the third letter of codons of an extreme thermophile. DNA sequence of the isopropylmalate dehydrogenase of Thermus thermophilus. J Biol Chem. 1984 Mar 10;259(5):2956–2960. [PubMed] [Google Scholar]
  8. Koyama Y., Hoshino T., Tomizuka N., Furukawa K. Genetic transformation of the extreme thermophile Thermus thermophilus and of other Thermus spp. J Bacteriol. 1986 Apr;166(1):338–340. doi: 10.1128/jb.166.1.338-340.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kunai K., Machida M., Matsuzawa H., Ohta T. Nucleotide sequence and characteristics of the gene for L-lactate dehydrogenase of Thermus caldophilus GK24 and the deduced amino-acid sequence of the enzyme. Eur J Biochem. 1986 Oct 15;160(2):433–440. doi: 10.1111/j.1432-1033.1986.tb09991.x. [DOI] [PubMed] [Google Scholar]
  10. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  11. Miles E. W., Kawasaki H., Ahmed S. A., Morita H., Morita H., Nagata S. The beta subunit of tryptophan synthase. Clarification of the roles of histidine 86, lysine 87, arginine 148, cysteine 170, and cysteine 230. J Biol Chem. 1989 Apr 15;264(11):6280–6287. [PubMed] [Google Scholar]
  12. Nagata S., Hyde C. C., Miles E. W. The alpha subunit of tryptophan synthase. Evidence that aspartic acid 60 is a catalytic residue and that the double alteration of residues 175 and 211 in a second-site revertant restores the proper geometry of the substrate binding site. J Biol Chem. 1989 Apr 15;264(11):6288–6296. [PubMed] [Google Scholar]
  13. Nichols B. P., Yanofsky C. Nucleotide sequences of trpA of Salmonella typhimurium and Escherichia coli: an evolutionary comparison. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5244–5248. doi: 10.1073/pnas.76.10.5244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nishiyama M., Matsubara N., Yamamoto K., Iijima S., Uozumi T., Beppu T. Nucleotide sequence of the malate dehydrogenase gene of Thermus flavus and its mutation directing an increase in enzyme activity. J Biol Chem. 1986 Oct 25;261(30):14178–14183. [PubMed] [Google Scholar]
  15. Sato S., Nakada Y., Kanaya S., Tanaka T. Molecular cloning and nucleotide sequence of Thermus thermophilus HB8 trpE and trpG. Biochim Biophys Acta. 1988 Sep 7;950(3):303–312. doi: 10.1016/0167-4781(88)90126-1. [DOI] [PubMed] [Google Scholar]
  16. Sato S., Nakazawa K., Shinomiya T. A DNA methylase from Thermus thermophilus HB8. J Biochem. 1980 Sep;88(3):737–747. doi: 10.1093/oxfordjournals.jbchem.a133026. [DOI] [PubMed] [Google Scholar]
  17. Singleton R., Jr, Amelunxen R. E. Proteins from thermophilic microorganisms. Bacteriol Rev. 1973 Sep;37(3):320–342. doi: 10.1128/br.37.3.320-342.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  19. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  20. Yutani K., Ogasahara K., Tsujita T., Kanemoto K., Matsumoto M., Tanaka S., Miyashita T., Matsushiro A., Sugino Y., Miles E. W. Tryptophan synthase alpha subunit glutamic acid 49 is essential for activity. Studies with 19 mutants at position 49. J Biol Chem. 1987 Oct 5;262(28):13429–13433. [PubMed] [Google Scholar]
  21. van Randen J., Venema G. Direct plasmid transfer from replica-plated E. coli colonies to competent B. subtilis cells. Identification of an E. coli clone carrying the hisH and tyrA genes of B. subtilis. Mol Gen Genet. 1984;195(1-2):57–61. doi: 10.1007/BF00332724. [DOI] [PubMed] [Google Scholar]

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